Butyrophilin is expressed in mammary epithelial cells from a single-sized messenger RNA as a type I membrane glycoprotein.
We investigated the expression of butyrophilin in eukaryotic cells with a view to determining the number of mRNA species, the incorporation of the peptide chain into microsomes, and the topology of the processed protein in biological membranes. Butyrophilin is synthesized from a single sized mRNA in both bovine and murine lactating mammary tissue and associates with microsomal membranes with a type I topology (Nexo.Ccyto) via a single hydrophobic anchor in the middle of the sequence. Several isoelectric variants of the protein were detected in cellular membranes from lactating bovine mammary tissue and in the milk-fat-globule membrane. We found no evidence for soluble forms of butyrophilin in postmicrosomal supernatants. The 66-kDa protein appears to be subjected to limited proteolysis, giving rise to a 62-kDa fragment lacking the C terminus and to other more minor fragments of lower Mr in the milk-fat-globule membrane. Antipeptide antibodies to epitopes within the N- and C-terminal domains were used to show that butyrophilin retains a type I topology in plasma membranes when expressed in insect cells from a baculovirus vector, and in secreted milk-fat globules. These data do not agree with previous suggestions that butyrophilin may exist in cytoplasmic soluble forms, or be reorganized in the plane of the lipid bilayer during secretion in lipid droplets from mammary cells. The results are discussed with reference to the role butyrophilin may play as the principal scaffold for the assembly of a complex with xanthine oxidase and other proteins that functions in the budding and release of milk-fat globules from the apical surface during lactation.
Banghart
L R
LR
Department of Animal and Avian Sciences, University of Maryland, College Park, Maryland 20742, USA.
Chamberlain
C W
CW
Velarde
J
J
Korobko
I V
IV
Ogg
S L
SL
Jack
L J
LJ
Vakharia
V N
VN
Mather
I H
IH
D000818:Animals; D000071797:Butyrophilins; D002417:Cattle; D003001:Cloning, Molecular; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000235:genetics; D015151:Immunoblotting; D008321:Mammary Glands, Animal / Q000737:chemistry*; D008562:Membrane Glycoproteins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D008894:Milk Proteins / Q000737:chemistry / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis; D014176:Protein Biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis*; D017421:Sequence Analysis; D018411:Spodoptera / Q000235:genetics
D000071797:Butyrophilins; D008562:Membrane Glycoproteins; D008894:Milk Proteins; D010446:Peptide Fragments; D012333:RNA, Messenger